【摘要】 目的:观察高压氧(hyperbaric oxygen,HBO)对肾脏缺血再灌注损伤的保护作用并探讨其作用机制。方法:56只SD大鼠被随机分为三组,假手术组(n=8);I/R组(n=24),夹闭双肾动脉45分钟后恢复血流灌注;I/R+HBO组(n=24),夹闭双肾动脉45分钟并在恢复血流后1 h、24 h、48 h行HBO治疗,每次HBO后采血并取双肾,比色法测定血浆尿素氮(BUN)、肌酐(Cr)值,原位末端标记(TUNEL)法检测肾小管上皮细胞凋亡情况,实时定量PCR法检测促凋亡基因Bax的mRNA含量。结果:与sham组(BUN值为9.563±1.384 mmol/L;Cr值为45.912±2.685 mmol/L,TUNEL值为2.088%)比较,I/R组大鼠再灌注1小时尿素氮(12.5±1.487 mmol/L)和血肌酐水平(51.388±3.092 mmol/L)升高,但差异无统计学意义,而TUNEL阳性细胞数(9.775%)和Bax的mR-NA(3.219±0.427)表达水平均显著升高(P<0.05),再灌注24小时及48小时后尿素氮(28.087±2.012 mmol/L、41.225±1.397mmol/L)和血肌酐(241.75±11.853 mmol/L、278.75±12.578 mmol/L)水平、TUNEL阳性细胞数(12.512%、14.413%)和Bax的mRNA(5.541±0.227、6.407±0.291)表达水平均显著升高(P<0.05);而HBO治疗可显著降低再灌注24小时及48小时的大鼠尿素氮(14.15±1.397 mmol/L、25.962±2.497 mmol/L)和血肌酐(146.375±8.782 mmol/L、210.125±11.519 mmol/L)水平(P<0.05),但仍显著高于假手术组(P<0.05)。结论:HBO治疗可以改善I/R后肾功能,其作用机制可能与在早期明显降低Bax的mRNA表达,减轻肾小管上皮细胞凋亡有关。更多还原

【Abstract】 Objective: To investigate the effect and mechanisms of hyperbaric oxygen(HBO) on the ischemia reperfusion injury in kidney.Methods: 56 SD rats were randomly divided into three groups: the sham group(n=8),I/R group(n=24) and I/R+HBO group(n=24).The I/R group group received bibateral kidney ischemia for 45 min and the I/R+HBO group received additional HBO therapy at the 1 h,24 h and 48 h after reperfusion.At the end of each HBO therapy,bilateral kidneys were got and renal function was determined by measuring the plasma blood urea nitrogen(BUN) and creatinine(Cr).Tubular cell apoptosis was confirmed by terminal deoxynucleotidyl transferase(TdT)-mediated dUTP-biotin nick end labeling(TUNEL) assay.The mRNA expression of Bax in kidney was detected by using Real-time RT-PCR.Results: Compared to the sham group(BUN: 9.563 ± 1.384 mmol/L;Cr: 45.912 ± 2.685 mmol/L,TUNEL 2.088 %),blood urea nitrogen(12.5 ±1.487 mmol/L) and serum creatinine levels(51.388±3.092 mmol/L) were increased in group I/R after 1 h of reperfusion,but no statistical difference was found,while the number of positive TUNEL cells(9.775 %) and Bax mRNA(3.219 ±0.427) expression level were significantly increased(P<0.05).Blood urea nitrogen(28.087±2.012 mmol/L for 24 h,41.225± 1.397 mmol/L for 48 h) and serum creatinine(241.75±11.853 mmol/L for 24 h,278.75 ± 12.578 mmol/L for 48 h) level,the number of TUNEL positive cells(12.512 % for 24 h,14.413% for 48 h) and Bax mRNA(5.541±0.227 for 24 h,6.407±0.291 for 48 h) levels were significantly increased(P<0.05) after 24 and 48 h of reperfusion.HBO treatment can significantly reduce rat blood urea nitrogen(14.15 ±1.397 mmol/L for 24 h,25.962 ±2.497 mmol/L for 48 h) and serum creatinine(146.375±8.782 mmol/L for 24 h,210.125±11.519 mmol/Lfor 48 h) level(P<0.05) after 24 h and 48 h of reperfusion,but still higher than that in sham operation group(P<0.05).Conclusion: HBO treatment can improve renal function induced by I/R,which may be caused by the reduction of Bax mRNA expression and the amelioration of renal tubular epithelial cell apoptosis in the early stages.更多还原